EFFECT OF TEMPERATURE ON THE WIDE ANGLE X-RAY DIFFRACTION OF NANOCRYSTALLINE

TRIACYLGLYCEROLS

Abstract

Many fat-structured based foods such as chocolate, coco butter and margarine are highly
influenced by the fat crystal network formed during the processing period. And x-ray diffraction
is widely used in measuring the nanocrystalline structure. However, the peak position of wide
angle x-ray diffraction patterns of nanocrystalline triacylglycerols is affected by the
temperature. To observe this effect quantitatively, pure triacylglycerol and its mixtures were
crystalized crystallized in a desired polymorph in capillaries or a Couette system. And the
crystallized samples were kept at different temperatures and shear rates. The detailed WAXD
patterns were obtained using in-house x-ray and synchrotron x-ray sources. When the
temperature increased, the d-spacing for the peaks with small d-spacing increased as well.
However, the d-spacing for other peaks remained unchanged or just had a very small change.
The relationship between differences of d-spacing for the peaks with small d-spacing and
temperature can be used to estimate the real sample temperature, especially under higher
shear rate. This research provides a new way to monitor and control the temperature of the
system under study and the effect of high shear rate on nanocrystalline triacylglycerol
crystallization.

1. INTRODUCTION

Fat-contained food are necessary part of our daily diet. The physical properties such as
sensation, flavor and text of food are derived from the network structure of the crystalline fat
(Narine and Marangoni, 1999,1 Wright et al., 2000, Mazzanti et al., 2005). These sensorial
attributes, such as texture, spread ability and mouth-feel, are dependent on the mechanical
strength of the fat crystal network (Narine and Marangoni, 1999).
Fats are mainly present as complex triglyceride, which consist the underlying nanoscale and
meso-scale crystalline structure. These unique constitute and structure make the fats become
an effective energy source compared with carbohydrates and proteins. And hydrophobic
property of fats allows it stack in the tissue cell a stabilized form without interrupting osmotic
balance of cell.
Fats are also necessary functionally mediator such as homone, vitamin and intercellular
messenger. However, the excessive intake of fats leads to dangerous heathy issues like clinically
obese, type- II diabetes and cardiovascular disease. The most effective solution is to change the
food present at consumer’s table, which means manipulate and replace potentially unhealthy
fats, which can be archived easily than the pharmaceutical intervention to improve the citizen’s
overall heath levels.
However, this put new challenge to food industry because the consumers are picky at sensory
of food. Thus, the fully understanding of edible fats are necessary to satisfy the demand of
palatable but heathy food. i Fat crystallization, the formation of solid fat crystals, always occurs
during the industrial manufacturing and the storage of the fat-based foods. It has a great
impact in the texture, shelf life and food quality of the fat-based foods (Metin and Hartel,
2005).
With further research, it’s possible to design and engineer rational food material and industrial
scale machine. Also, the new researches in fats can be used to optimize and improve
manufacturing process with appeling benefits. A comprehensive understanding of the fat
crystallization process of dietary fat from physics, chemistry and biochemistry is needed (Sato
et al., 1999).
Many factors, like solid fat content, microstructure of the crystal and the types of polymorph
have great influence in determining the crystallized fat structure. In addition, the processing
conditions, such as crystallization temperatures, shear rates and cooling rates are closely
related to fat crystallization.
The complexity of natural fats in the fat-based foods limits the accuracy of manufacturing and
results in uncertain quality of the products. In this research, the main aim is to observe the
behaviour of nanostructure during the lipid crystallization process under several constant
temperatures and shear rates using x-ray diffraction techniques. It was noticed in previous
studies that the peak position of wide angle x-ray diffraction (WAXD) patterns is affected by the
temperature (Mazzanti, unpublished). In order to observe this effect quantitatively,
crystallization of different kinds of triacylglycerols (TAGs) and their mixtures, which are
normally the main or critical components of the food products, are studied at different
controlled temperatures and shear flow.
Wide angle x-ray diffraction (WAXD or WAXS) is a technique used to determine the crystal
structure, crystallinity and molecular orientation in crystal. The WAXD reflects the patterns of
the lateral distance between two hydrocarbon chains in crystal, which can be used to identify
the different polymorphic forms of TAGs. Different WAXD peaks change their position as the
temperature changes and this provides a new attempt to study the change of sample
temperature when a shear rate is applied.
Shear is an regular industrial manufacturing process widely used in the food industry to obtain
the homogeneous products. Shears flow can produce the viscous heat which can alter expected
property, such as delay of onset of crystallization, change in solid fat content (SFC), modification
of the composition of solid and liquid phase and induction of phase transition, based on the
enhancement of heat transfer from thermodynamic prospective. And the anisotropy of the
nano-crystal is also evident by these changes. The findings of this research should help future
studies of academic researchers and the food industry to develop a deeper knowledge of lipid
crystallization, leading to better products and manufacturing procedures.

The aim of this study is to investigate the characteristics of the triacylglycerol crystallizing under
different temperatures and shear rates. The pure and mixed triacylglycerol crystal were used to
study the influence of various temperature on different WAXD peaks. In addition, the different
shear rates were set up to investigate the influence on the WAXD peaks. A summary was made
based on the relation between WAXD peaks and temperature, which can be used as an intrinsic
thermometer applied in the study of thermo-mechanical property of triacylglycerol crystals
under high shear rates.
2. Experimental

2.1 Materials
Trilaurin (LLL), trimyristin (MMM), tripalmitin (PPP) and tristearin (SSS) were
purchased from Sigma-Aldrich Chemical Co. and had a purity of at least 99%. No
further purification was carried out.
The binary mixtures (dry blends) were prepared by mixing the weighted samples in
composition on a weight per weight basis. For example, 3L7M represents the ratio of
LLL and MMM is 30:70 on weight base. 3L7M, 5L5M, 7L3M, 3M7P, 5M5P, 7M3P,
3P7S, 5P5S, 7P3S and 5L5S were prepared.

2.1.1 X-ray capillary

The samples mentioned above were melted at approximately 100 °C using a hot
plate (Cole-Parmer, USA). 20-30 μL of sample was transferred to the x-ray capillary
(Charles Supper Co., 1.5 mm diameter, 10 μm wall) by using a preheated disposable
capillary tube with a wire plunger (Drummond Scientific Company, Wiretrol® II, Cat.
Number 5-000- 2010, 5 μL and l0 μL) and sealed by fire.
Cocoa butter and the commercial product, 99% dark chocolate, were also used in
this research. Cocoa butter was stored at -20 °C in the fridge for a long time and it
was crystallized in the β form. 99% dark chocolate was purchased in the local
supermarket and the chocolate was crystallized in β form (V phase). The solid cocoa
butter and 99% dark chocolate were chopped into tiny pieces and put into the
capillaries without melting and sealed by fire.

2.1.2 Mini Couette cell

6B4P and 6B4S were prepared as samples. BBB was used as a solvent to dissolve PPP
and SSS according to the required ratio in composition on a weight per weight basis.
The BBB: PPP ratio and the BBB: SSS ratio was 60:40. The samples were melted and
placed in the sample tubes, which fitted the mini Couette cell.

2.2 In-house wide angle X-Ray diffraction measurements

Figure 1.1 In-house XRD set up.

The in-house WAXD experimental set up is shown in Figure 1- 1. The x-ray generator had a
Commercial GeniX x-ray source (Xenocs Corporation, Sassenage, France), which produced
0.7093 Å wavelength Mo Kα radiation.
The x-rays were focused by a mirror and then collimated by two sets of scatterless slits which
were aligned on a small diameter vacuum fly-path. The x-ray beam was projected to the sample
capillary right after it passed the fly-path. The beam size at the sample was approximately 0.4
mm x 0.5 mm.
The detector used in our in-house x-ray experimental set up was XRI-UNO/Si 2D x-ray detector
(XRAY-IMATEK, Barcelona, Spain). The detector was built as a single chip array of silicon sensors
and its active area was 14 mm × 14 mm. The pixel size (ps) was 0.055 mm ×0.055 mm. The
detector camera was controlled and all the images were collected by software named ‘XRI-
UNO’. There was an enclosure around the whole x-ray system to isolate the surroundings and
minimize the danger of x-ray exposure.

2.3
Marangoni, Alejandro G. Acevedo, Nuria Maleky, Fatemeh Co, Edmund Peyronel,
i

Fernanda Mazzanti, Gianfranco Quinn, Bonnie Pink, David

Structure and functionality of edible fats